title: Accurate detection of upper tract urothelial carcinoma in tissue and urine by means of quantitative GDF15, TMEFF2 and VIM promoter methylation.
abstract: OBJECTIVE
  Upper tract urothelial carcinoma (UTUC) accounts for 5-10% of all urothelial tumours. It is mostly diagnosed at advanced stages, entailing a worse prognosis, owing to the lack of early and specific symptoms as well as of effective diagnostic tools. We previously identified a panel of epigenetic biomarkers (GDF15, TMEFF2 and VIM promoter methylation) that accurately identifies bladder cancer in urine. Herein, we assessed the performance of the same panel for UTUC detection and prognosis, in tissue and urine.
  
  
  METHODS
  Methylation levels of reference and target genes were determined using real-time quantitative methylation-specific polymerase chain reaction (MSP) in bisulphite-modified DNA of 57 UTUC tissues, 36 normal upper tract urothelium (NUTUs), 22 urines from UTUC suspects and 20 urines from controls. Receiver operator characteristics (ROC)-curve analysis was performed to determine the performance of the biomarker panel and survival analyses were conducted to evaluate their prognostic value.
  
  
  RESULTS
  Methylation levels of GDF15, TMEFF2 and VIM were significantly higher in UTUC compared to NUTUs (P=0.022; P<0.001; P<0.001, respectively). The panel accurately identified UTUC with 100% and 91% sensitivity, corresponding to an area under the curve of 1.000 and 0.923 in tissue and urines, respectively, with 100% specificity. Low VIM promoter methylation levels independently predicted poor disease-specific survival.
  
  
  CONCLUSIONS
  GDF15, TMEFF2 and VIM promoter methylation allows for accurate identification of UTUC, in tissue and urine and VIM methylation provides relevant prognostic information, especially in high-stage disease. This assay may improve the clinical management of UTUC patients.
authors.affiliation: Cancer Epigenetics Group, Research Center of the Portuguese Oncology Institute - Porto, Rua Dr. António Bernardino de Almeida, 4200-072 Porto, Portugal; Department of Genetics, Portuguese Oncology Institute - Porto, Rua Dr. António Bernardino de Almeida, 4200-072 Porto, Portugal.
authors.forename: Sara
authors.initials: S
authors.lastname: Monteiro-Reis
authors.affiliation: Department of Pathology, Portuguese Oncology Institute - Porto, Rua Dr. António Bernardino de Almeida, 4200-072 Porto, Portugal.
authors.forename: Luís
authors.initials: L
authors.lastname: Leça
authors.affiliation: Cancer Epigenetics Group, Research Center of the Portuguese Oncology Institute - Porto, Rua Dr. António Bernardino de Almeida, 4200-072 Porto, Portugal; Department of Genetics, Portuguese Oncology Institute - Porto, Rua Dr. António Bernardino de Almeida, 4200-072 Porto, Portugal.
authors.forename: Mafalda
authors.initials: M
authors.lastname: Almeida
authors.affiliation: Department of Epidemiology, Portuguese Oncology Institute - Porto, Rua Dr. António Bernardino de Almeida, 4200-072 Porto, Portugal.
authors.forename: Luís
authors.initials: L
authors.lastname: Antunes
authors.affiliation: Department of Pathology, Portuguese Oncology Institute - Porto, Rua Dr. António Bernardino de Almeida, 4200-072 Porto, Portugal.
authors.forename: Paula
authors.initials: P
authors.lastname: Monteiro
authors.affiliation: Department of Pathology, Portuguese Oncology Institute - Porto, Rua Dr. António Bernardino de Almeida, 4200-072 Porto, Portugal.
authors.forename: Paula C
authors.initials: PC
authors.lastname: Dias
authors.affiliation: Department of Urology, Portuguese Oncology Institute - Porto, Rua Dr. António Bernardino de Almeida, 4200-072 Porto, Portugal.
authors.forename: António
authors.initials: A
authors.lastname: Morais
authors.affiliation: Department of Urology, Portuguese Oncology Institute - Porto, Rua Dr. António Bernardino de Almeida, 4200-072 Porto, Portugal.
authors.forename: Jorge
authors.initials: J
authors.lastname: Oliveira
authors.affiliation: Cancer Epigenetics Group, Research Center of the Portuguese Oncology Institute - Porto, Rua Dr. António Bernardino de Almeida, 4200-072 Porto, Portugal; Department of Pathology, Portuguese Oncology Institute - Porto, Rua Dr. António Bernardino de Almeida, 4200-072 Porto, Portugal; Department of Pathology and Molecular Immunology, Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto, Rua de Jorge Viterbo Ferreira n° 228, 4050-313 Porto, Portugal.
authors.forename: Rui
authors.initials: R
authors.lastname: Henrique
authors.affiliation: Cancer Epigenetics Group, Research Center of the Portuguese Oncology Institute - Porto, Rua Dr. António Bernardino de Almeida, 4200-072 Porto, Portugal; Department of Genetics, Portuguese Oncology Institute - Porto, Rua Dr. António Bernardino de Almeida, 4200-072 Porto, Portugal; Department of Pathology and Molecular Immunology, Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto, Rua de Jorge Viterbo Ferreira n° 228, 4050-313 Porto, Portugal. Electronic address: carmenjeronimo@ipoporto.min-saude.pt.
authors.forename: Carmen
authors.initials: C
authors.lastname: Jerónimo
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism / Q000652:urine*; D019175:DNA Methylation; D005260:Female; D055436:Growth Differentiation Factor 15 / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D008875:Middle Aged; D009363:Neoplasm Proteins / Q000235:genetics* / Q000378:metabolism; D011379:Prognosis; D011401:Promoter Regions, Genetic; D001749:Urinary Bladder Neoplasms / Q000235:genetics* / Q000473:pathology / Q000652:urine*; D019459:Urothelium / Q000378:metabolism / Q000473:pathology*
chemical_list: D014408:Biomarkers, Tumor; C526140:GDF15 protein, human; D055436:Growth Differentiation Factor 15; D008565:Membrane Proteins; D009363:Neoplasm Proteins; C423567:TMEFF2 protein, human
keywords: Detection; Methylation; Upper tract urothelial cancer; Urine markers